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Caracterización Molecular Y Biológica Del Virus De La Rabia Que Circula En Revista Mexicana de Ciencias Pecuarias ISSN: 2007-1124 [email protected] Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias México Loza-Rubio, Elizabeth; Nadin-Davis, Susan A.; Morales Salinas, Elizabeth Caracterización molecular y biológica del virus de la Rabia que circula en zorrillos de México enfocado a la variante del gen de la fosfoproteína (P) Revista Mexicana de Ciencias Pecuarias, vol. 3, núm. 2, abril-junio, 2012, pp. 155-170 Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias Morelos, México Disponible en: http://www.redalyc.org/articulo.oa?id=265624467002 Cómo citar el artículo Número completo Sistema de Información Científica Más información del artículo Red de Revistas Científicas de América Latina, el Caribe, España y Portugal Página de la revista en redalyc.org Proyecto académico sin fines de lucro, desarrollado bajo la iniciativa de acceso abierto CARACTERIZACIÓN MOLECULAR Y BIOLÓGICA DEL VIRUS DE LARev RABIA Mex CiencEN ZORRILLOS Pecu 2012;3(2):155-170 Caracterización molecular y biológica del virus de la Rabia que circula en zorrillos de México enfocado a la variante del gen de la fosfoproteína (P) Molecular and biological properties of rabies viruses circulating in Mexican skunks: focus on P gene variation Elizabeth Loza-Rubioa, Susan A. Nadin-Davisb, Elizabeth Morales Salinasc RESUMEN El objetivo de este estudio fue la caracterización molecular y biológica del virus de la rabia en zorrillos de México, comparando una porción de la secuencia viral del gen P, con regiones correspondientes de variantes de virus rábico adaptadas en zorrillos y pertenecientes al genotipo 1 (virus de la rabia “clásica”), que circulan en América. Como parte de la caracterización biológica se incluyó el periodo de incubación y las lesiones histopatológicas después de la inoculación del virus por vía intracerebral en ratones. De acuerdo a los resultados de estudios filogenéticos, las cepas de zorrillo mexicano (variante antigénica (AV8 y AV10) son muy distintas en cuanto a su evolución. El aislado AV10 de Baja California Sur está muy relacionado con los virus que circulan en zorrillos de California; mientras que el aislado AV8 de San Luis Potosí tuvo mayor relación con la cepa del zorrillo del Centro/Sur que circula en los estados del sur, como Texas. Estas variantes se reflejaron en algunas propiedades biológicas de ambas cepas en ratones. PALABRAS CLAVE: Rabia zorrillos, Epidemiología molecular, Periodo de incubación, Gene P. ABSTRACT The objective of this work was to characterize molecularly rabies viruses from Mexican skunks, by comparison of a portion of the viral P gene sequence with corresponding regions from other skunk-adapted rabies virus variants and with other genotype 1 rabies viruses that circulate in the Americas. Furthermore, incubation period and histopathologic lesions after virus inoculation by the intra-cerebral route in mice was characterized. According to the results of phylogenetic studies the Mexican skunk strains (Antigenic Variant (AV) 8 and AV10) are evolutionarily quite distinct. The AV10 isolate from South Baja California is quite closely related to the viruses that circulate in Californian skunks; while the AV8 isolate from San Luis Potosí was most closely related to the South central skunk strain that circulates in southern states such as Texas. These variations were reflected in some biological properties of both strains in mice. KEY WORDS: Rabies, Skunks, Molecular epidemiology, Incubation period, P gen. INTRODUCCIÓN INTRODUCTION La rabia es una enfermedad en la que se presenta The disease of rabies is an acute, progressive, encefalitis aguda, progresiva e incurable, causada incurable encephalitis caused by all members of the por todos los miembros del género Lyssavirus de la Lyssavirus genus of the Rhabdoviridae family. The familia Rhabdoviridae. Este género contiene hasta genus Lyssavirus contains 11 viruses: rabies virus Recibido el 4 de mayo de 2011. Aceptado el 28 de junio de 2011. a Centro Nacional de Investigaciones Disciplinarias en Microbiología Animal, INIFAP. Carretera México Toluca Km. 15.5, Colonia Palo Alto. CP 05110. México. [email protected]. Correspondencia al primer autor. b Rabies Center of Expertise, Ottawa Laboratories. Fallowfield, Canadian Food Inspection Agency (CFIA). Ottawa, Ont. Canadá K2H 8P9. c Departamento de Patología. FMVZ-UNAM. 155 Elizabeth Loza-Rubio, et al. / Rev Mex Cienc Pecu 2012;3(2):155-170 ahora 12 virus: el de la rabia (VRAB), virus del (RABV), Lagos bat virus, Mokola virus, Duvenhage murciélago Lagos, virus Mokola, virus Duvenhage, virus, European bat lyssaviruses types 1 and 2 Lyssavirus del murciélago europeo tipo 1 y 2 (EBLV- (EBLV-1 and EBLV-2), Australian bat lyssavirus, 1 y EBLV-2), Lyssavirus del murciélago Australiano, Aravan virus (ARAV), Khujand virus (KHUV), Irkut virus Aravan, virus Khujand, virus Irkut y virus virus, and West Caucasian bat virus(1). A proposed caucásico del oeste(1). Una nueva especie propuesta new species, Shimoni bat virus has recently been es el virus del murciélago de Shimoni que ha sido isolated from Hipposideros commersoni leaf-nosed recientemente aislado del murciélago con nariz en bats(2). Although several Lyssavirus genotypes are forma de hoja (Hipposideros commersoni)(2). now recognized world-wide(2,3) only genotype 1 Aunque varios genotipos de Lyssavirus son viruses, represented by classical serotype 1 rabies reconocidos mundialmente(2,3), sólo el virus virus, is known to circulate on the American genotipo 1, representado por el virus de la rabia Continent(4,5). serotipo 1 clásico, es sabido que circula en el continente Americano(4,5). In Mexico, urban rabies has been significantly reduced over the past decade due to intensive dog La rabia urbana en México ha sido control and dog vaccination campaigns(6). However, significativamente reducida durante la última década with the reduction of dog rabies it has become por control canino intensivo y campańas de increasingly evident that sylvatic rabies circulates vacunación(6). Sin embargo, al reducir el número in several wildlife species of both terrestrial and de casos de rabia en perros, ha sido muy notoria aerial life-styles(7,8,9,10). The vampire bat la presencia de rabia en varias especies silvestres (Desmodus rotundus) represents the host that most tanto de vida terrestre como aérea(7,8,9,10). El frequently transmits rabies to humans and livestock, murciélago vampiro (Desmodus rotundus) representa especially cattle, but the potential role of skunks in al portador que con mayor frecuencia transmite la causing human disease has been documented(11). rabia a humanos y al ganado, principalmente bovino, The principal species of skunk that appears to act pero también se ha informado del potencial que as a rabies reservoir in Mexico is the spotted skunk presentan los zorrillos para causar esta enfermedad (Spilogale putorius) although other species such as en humanos(11). La especie principal de zorrillo the hog-nosed skunk (Conepatus leuconotus) have que actúa como reservorio de rabia en México es been reported as rabid(11). In contrast, in Canada el zorrillo moteado (Spilogale putorius), aunque and the US the species of skunk most usually otras especies como el zorrillo de nariz porcina associated with rabies is the striped skunk (Mephitis (Conepatus leuconotus) han sido documentadas mephitis)(12). como rabiosas(11). Por otro lado, en Canadá y en Estados Unidos de América la especie de zorrillo Antigenic methods, employing a small panel of más comúnmente asociado con la rabia es el zorrillo monoclonal antibodies, have been found to identify rayado (Mephitis mephitis)(12). the following antigenic variants (AVs) in Mexican wildlife reservoirs: AV3 and 11 carried by vampire Se ha encontrado que los métodos antigénicos, que bats, AV4 and 9 in Mexican free-tailed bats utilizan un panel reducido de anticuerpos (Tadarida brasiliensis), AV7 recovered from bobcats monoclonales, identifican las siguientes variantes and foxes and two antigenic types, AV8 and AV10 antigénicas (AV) en reservorios silvestres mexicanos: associated with skunks in two different parts of the AV3 y 11 cuyos portadores son los murciélagos country, a central region including San Luis Potosi vampiros, AV4 y 9 detectadas en murciélagos de (SLP) state and South Baja California (SBC) cola de ratón (Tadarida brasiliensis), AV7 de linces respectively(9). It is known that for example AV 8 and y zorros, y dos tipos antigénicos, AV8 y AV10 AV 10 refer to variants of rabies virus maintained in asociados con zorrillos en dos partes del país, una skunk populations, and so on. It is known that for región central que incluye a San Luis Potosí (SLP) example AV 8 and AV 10 refer to variants of rabies y Baja California Sur (BCS), respectivamente(9). virus maintained in skunks populations, and so on. 156 CARACTERIZACIÓN MOLECULAR Y BIOLÓGICA DEL VIRUS DE LA RABIA EN ZORRILLOS Es sabido por ejemplo que las variantes AV8 y Partial characterization of rabies virus N gene AV10 se refieren a variantes antigénicas del virus sequences of some representatives of these variants de la rabia mantenidas en poblaciones de zorrillos. did not associate the Mexican SBC skunk strain together with any other rabies variant while the La caracterización parcial de las secuencias del gen bobcat isolates appeared to be closely related to fox N del virus de la rabia de algunas representantes strains of the southern US(10,13). In addition, genetic de estas variantes no asoció la cepa de zorrillo characterisation of the viral G/G-L region has mexicano de BCS con ninguna variante de rabia, facilitated development of rapid genetic tests employing mientras que los aislados del lince mostraron estar restriction fragment length polymorphisms (RFLPs) muy relacionados con cepas de zorros del sur de to discriminate dog and vampire bat rabies isolates Estados Unidos de América(10,13). Adicionalmente, and also identified a discrete pattern described as la caracterización genética de la región G/G-L viral “hypervariable” that was associated with viruses ha facilitado el desarrollo de pruebas genéticas from skunks(14).
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